The present invention provides sequences capable of inhibiting osteopontin (OPN) expression. In particular, the sequences provided herein are antisense osteopontin oligonucleotide sequences. The present invention further relates to methods for treating restenosis using antisense osteopontin oligonucleotide sequences, and in particular, to treating restenosis following vascular surgery.
Atherosclerosis (for review see Ross, R. (1993) Nature 362:801-809 and Hajjar et al., (1995) Amer. Scientist 83:460-467) is the principal cause of heart attacks, stroke, gangrene and loss of function of extremities. It accounts for approximately 50% of all mortalities in the USA, Europe and Japan (Ross, R. (1993) Nature 362:801-809). The present therapeutic strategies for severe atherosclerosis in coronary arteries rely on angioplasty procedures (e.g., percutaneous trans-luminal coronary angioplasty (PTCA), directional coronary atherectomy (DCA) or related angioplasty procedures), and coronary artery bypass surgery. For example, PTCA is the primary treatment modality in many patients with coronary heart disease. PTCA can relieve myocardial ischemia in patients with coronary artery disease by reducing lumen obstruction and improving coronary bloodflow.
While the use of interventional procedures has grown rapidly, reocclusion (or restenosis) of arteries is a serious complication which occurs in 30-50% of patients undergoing various angioplasty procedures within 3 days to 3 months. Restenosis results in significant morbidity and mortality and frequently necessitates further interventions, such as repeat angioplasty or coronary bypass surgery.
Although the processes responsible for restenosis are not completely understood, restenosis has been suggested to occur. at least in part, as a result of local inflammation, thrombosis and smooth muscle cell migration (Ferrell et al. (1992) Circulation 85:1630-1631) and proliferation (Austin et al. (1985) J. Am. Coll. Cardiol. 6:369-375; Giraldo et al. (1985) Arch. Pathol. Lab. Med. 109:173-175) within the intima of coronary arteries. To date, no post-angioplasty treatment has proven effective in the prevention or treatment of restenosis.
Thus, there is a need for methods and compositions for preventing and/or treating restenosis. Preferably, these methods and compositions are specific in their effect, easy to administer, and are effective over a short period of administration with minimal adverse side-effects.
The present invention discloses novel osteopontin antisense sequences which are useful for the treatment and prevention of restenosis. The present invention further discloses methods of diminishing osteopontin expression in a subject capable of developing restenosis in a tissue, methods of treating restenosis in a subject suspected of being capable of developing restenosis in a tissue, methods of reducing osteopontin expression in a subject undergoing angioplasty, methods of treating restenosis in a subject undergoing angioplasty, and methods of detecting restenosis in a subject.
In particular, the invention provides an antisense sequence comprising a nucleic acid sequence complementary to at least a portion of the human osteopontin cDNA polynucleotide listed herein as SEQ ID NO:15. While it is not intended that the present invention be limited to any particular antisense sequence, in one preferred embodiment the antisense sequence is selected from the group consisting of SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, and SEQ ID NO:13. In addition, though the present invention is not limited to a particular type of linkage, in a more preferred embodiment, the antisense sequence comprises one or more phosphorothioate linkages. In a yet more preferred embodiment, the antisense sequence is entrapped in a liposome.
The present invention also provides pharmaceutical compositions comprising a pharmaceutically acceptable excipient and an antisense sequence comprising a nucleic acid sequence complementary to at least a portion of the polynucleotide of SEQ ID NO:15.
Further provided by the instant invention are methods of diminishing osteopontin expression, comprising: a) providing: i) a subject suspected of being capable of developing restenosis in a tissue; and ii) an osteopontin antisense sequence complementary of at least a portion of the polynucleotide of SEQ ID NO:15; and b) administering an amount of the sequence to the subject under conditions such that the osteopontin expression is diminished.
Without intending to limit the present invention to any particular subject, in one embodiment, the subject is undergoing angioplasty. Also without limiting the invention to a particular surgical method, in a more preferred embodiment, the angioplasty is selected from the group consisting of percutaneous trans-luminal coronary angioplasty and directional coronary atherectomy.
In an alternative embodiment, and without limiting the invention to a particular type of tissue, the tissue is coronary vascular tissue. In a preferred embodiment, the coronary vascular tissue is arterial.
In yet another alternative embodiment, without intending to limit the invention to a particular sequence, the osteopontin antisense sequence is selected from the group consisting of SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12 and SEQ ID NO:13.
Although it is not intended that the present invention be limited to a particular method of administering, in a further alternative embodiment, the administering is parenteral. In a preferred embodiment, the parenteral administering is intraarterial (i.e., to the artery which is subjected to angioplasty). In yet a more preferred embodiment, the intraarterial administering is by using a catheter. In a particularly preferred embodiment, the catheter is a double balloon catheter.
In yet another alternative embodiment, the osteopontin antisense sequence is entrapped in a liposome.
The instant invention further provides methods of treating restenosis, comprising: a) providing: i) a subject suspected of being capable of developing restenosis in a tissue; and ii) an osteopontin antisense sequence complementary to at least a portion of the polynucleotide of SEQ ID NO:15; and b) administering an amount of the sequence to the subject under conditions such that the restenosis is diminished.
The present invention further provides methods of reducing osteopontin expression in a subject undergoing angioplasty, comprising: a) providing: i) a subject undergoing angioplasty; and ii) an osteopontin antisense sequence complementary to at least a portion of the polynucleotide of SEQ ID NO:15; and b) administering an amount of the sequence to the subject under conditions such that osteopontin expression is diminished. In one embodiment, and without intending to limit the invention to a particular sequence, the osteopontin antisense sequence is selected from the group consisting of SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12 and SEQ ID NO:13. In a preferred embodiment, the osteopontin antisense sequence comprises one or more phosphorothioate linkages. In a more preferred embodiment, the osteopontin antisense sequence is entrapped in a liposome. In yet a more preferred embodiment, the administering is substantially contemporaneous with the angioplasty. In a particularly preferred embodiment, the administering is by using a catheter. In a most preferred embodiment, the catheter is a double balloon catheter.
The present invention also provides methods of treating restenosis in a subject undergoing angioplasty, comprising: a) providing: i) a subject undergoing angioplasty; and ii) an osteopontin antisense sequence complementary of at least a portion of the polynucleotide of SEQ ID NO:15; and b) administering an amount of the sequence to the subject under conditions such that restenosis is diminished.
Also provided by the present invention are methods of detecting restenosis in a first subject, comprising detecting a higher level of osteopontin in a first tissue of a first subject suspected of being capable of developing restenosis in a second tissue relative to a level of osteopontin in said first tissue of a second subject substantially free of restenosis in said second tissue. In one embodiment, the first tissue is selected from the group consisting of blood and plasma. In a preferred embodiment, the first tissue comprises monocytes comprising the osteopontin. In a more preferred embodiment, the second tissue is coronary vascular tissue. In yet a more preferred embodiment, the coronary vascular tissue is arterial.